GSM6589714: PDAC IgG scRNA; Mus musculus; RNA-Seq
PDAC tumors were dissected out at d21 after tumor implantation and subsequently minced with razor blades in a cell culture dish before being digested with dissociation buffer (10x dissociation buffer: 40ml RPMI/DMEM with 1% Pen/Strep + 1mM NaPyr + 25mM HEPES + 400mg Collagenase  IV + 400mg Soybean Trypsin Inhibitor + 50mg Dispase II + 20mg DNAse) for 30 min at 37°C. Samples were then strained through a 70µM cell strainer, spun down at 420rcf and 4°C for 4 min. 4 tumors each per treatment condition were pooled and stained for sorting. The following cell populations were sorted into RPMI supplemented with 20% fetal bovine serum: Thy1.2+CD45.2+, Thy1.2-CD45.2+, YFP+CD45.2-. Sorted cells were spun down at 420 rcf at 4°C for 4 minutes, resuspended in PBS + 2% fetal bovine serum and mixed at 8k Thy1.2+CD45.2+, 10k Thy1.2-CD45.2+, 2k YFP+CD45.2- for a total of 20k cells per sample. Single cell RNA libraries were generated following the manufacturer's instructions of the Chromium Next GEM Single Cell 3' Reagent Kit v3.1 (10X Genomics).